Project:Plant species testing
Revision as of 21:01, 17 July 2012 by Mycoplasma (talk | contribs)
We are using primers for plants to test and refine our DNA extraction / PCR technique. First project is to test for peas. Inspiration in this paper.
First extraction and PCR - 17/7/12
We used two methods of extraction. Method 1:
- Blend peas with salt and detergent
- Incubate for 15 mins at 60 C (optional step not taken)
- Filter and centrifuge - recover supernatant
- Add proteinease K (0.8 ul - incubate for 1 hr at 50C then 94 C for 10 mins (inactivation of proteinase K)
- Precipitate with rubbing alcohol and meat tenderiser as an enzyme
- Centrifuge and remove supernatant to leave pellet
Method 2:
- 'Homogenize' with a pestle in 5% chelex solution for 1 min
- Vortex for 10s then boil for 5 mins, then vortex again for 10s
- Centrifuge for 1 min - recover supernatant and use as template